FIGS. 1A-1C. Solvent exposure of sidechains of framework residues in KOL and J539 Fvs and the residues which occur most frequently at these positions in the various human VH subgroups.
FIGS. 2A and 2B. Solvent exposure of sidechains of framework residues in KOL VL and the residues which occur most frequently at these positions in the various human V-lambda subgroups.
FIGS. 3A and 3B. Solvent exposure of sidechains of framework residues in J539 VL and the residues which occur most frequently at these positions in the various human V-kappa subgroups.
FIG. 4. Primers used to isolate DNA encoding murine kappa light chain variable region and murine IgG2a heavy chain variable region using PCR. Oligodeoxynucleotides used as PCR primers to generate a shortened IgG4 heavy chain. Oligodeoxynucleotides used in PCR to re-engineer the thymidine kinase (TK) promoter to facilitate the expression of the neomycin resistance gene. Oligodeoxynucleotide primers used in PCR to clone the IgH enhancer sequence. Oligodeoxynucleotides used as PCR primers to generate a human kappa light chain constant region.
FIG. 5. Oligodeoxynucleotides used in the construction of the xe2x80x9cveneeredxe2x80x9d 1B4 heavy and light chain variable regions plus those necessary to fuse the human signal and intronic sequenceds onto these variable regions.
FIG. 6. PCR-recombination strategy used in the veneering of the 1B4 kappa light chain variable region.
FIGS. 7A-7C. Outline of the insertion of the xe2x80x9cveneeredxe2x80x9d kappa light chain variable region and kappa constant region into the light chain expression vector.
FIG. 8. PCR-recombination strategy used in the veneering of the 1B4 heavy chain variable region.
FIGS. 9A an 9B. Outline of the insertion of the xe2x80x9cveneeredxe2x80x9d heavy chain variable region into the heavy chain expression vector.
FIGS. 10A and 10B. Outline of the insertion of neomycin selectable expression vector.
FIGS. 11A-10C. Outline of the insertion of hygromycin selectable expression vector.
FIG. 12. Amino acid sequence compleition of the xe2x80x9cveneeredxe2x80x9d-1B4, murine 1B4 and human Gal heavy chain variable regions and the xe2x80x9cveneeredxe2x80x9d 1B4, murine 1B4 and human Len kappa light chain variable regions. Check marks idicate the individual amino acid residues converted.
FIG. 13. Competitive binding assay of native murine 1B4 (open diamonds) and recombinant xe2x80x9cveneeredxe2x80x9d 1B4 (closed diamonds).
FIG. 14. Inmunogenicity of a veneered murine antibody in Rhesus monkeys is shown.